High-performance liquid chromatography of acid-stable and acid-labile phosphoamino acids.
A high-performance liquid chromatographic system has been developed which permits the separation of both acid-stable and acid-labile phosphoamino acids. An anion-exchange resin and two buffers of different ionic strength and near neutral pH are used. A low-ionic-strength buffer is used for the separation of N-omega-phosphoarginine and N-epsilon-phospholysine, while the higher-ionic-strength buffer permits the clear separation of tau-phosphohistidine, omicron-phosphoserine and omicron-phosphothreonine. An in-stream fluorometric detection system using omicron-phthalaldehyde permits the rapid analysis of samples containing as little as 25 pmoles of phosphoamino acid. This method has been applied to the detection of tau-phosphohistidine from alkaline digests of chemically phosphorylated calf thymus histone 4 and bovine myelin basic protein.
D000596:Amino Acids / Q000032:analysis*; D001120:Arginine / Q000031:analogs & derivatives / Q000032:analysis; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006639:Histidine / Q000031:analogs & derivatives / Q000032:analysis; D008239:Lysine / Q000031:analogs & derivatives* / Q000032:analysis; D009943:Organophosphorus Compounds / Q000032:analysis; D010768:Phosphoserine / Q000032:analysis; D010769:Phosphothreonine / Q000032:analysis
D000596:Amino Acids; D009943:Organophosphorus Compounds; D010769:Phosphothreonine; C015441:phospho-L-arginine; C025308:N-epsilon phospholysine; D010768:Phosphoserine; D006639:Histidine; D001120:Arginine; D008239:Lysine; C005436:phosphohistidine